Polymerization of G-actin by hydrodynamic shear stresses.
In the absence of Ca2+ G-actin can be polymerized by the application of shear stress in low ionic strength buffer. When G-actin in low ionic strength buffer containing EGTA was sheared for predetermined times under different velocity gradients, viscosity attained a maximal value, comparable to that obtained by seeding with F-actin nuclei, at a velocity gradient of 3000 s-1 after about one hour. Such flow-polymerized actin was indistinguishable from KCl-polymerized actin. Under similar conditions, EDTA which can bind both Ca2+ and Mg2+, gave a smaller effect than the Ca2+-chelating agent EGTA which binds Mg2+ weakly. When an Mg2+ salt was added to EDTA- or EGTA-containing buffer to give a free Mg2+ concentration of a few micromoles/liter, flow-induced polymerization was significantly enhanced. It appears that occupancy of only a small fraction of the high affinity binding sites by Ca2+ prevents flow-polymerization while Mg2+ may enhance this type of polymerization by replacing Ca2+. We speculate that the shear stress induces polymerization by promoting nucleation and that Ca2+ bound to the high affinity divalent cation binding site inhibits formation of the nuclei.
D000199:Actins; D001665:Binding Sites; D002118:Calcium; D004492:Edetic Acid; D004533:Egtazic Acid; D046911:Macromolecular Substances; D008274:Magnesium; D009994:Osmolar Concentration; D013314:Stress, Mechanical; D014783:Viscosity
D000199:Actins; D046911:Macromolecular Substances; D004533:Egtazic Acid; D004492:Edetic Acid; D008274:Magnesium; D002118:Calcium